Composition of matter and process for the control of drug resistant microorganisms



United States Patent Ofitice Bfilihld Patented Jan. 29, 1963 3,o75,s7a conrrosrriors or Marten Ann non CQN'ERQL RESETANT MEQRQ- (liRGANlSP/lfi Leonard Vincent Strully, 8-85 Plymouth thrive, Fairlawn, NJ. No Drawing. Filed Mar. El, 19m, Ser. N 16,188

19 Claims. (Q5. 167-55) I This invention relates to new compositions of matter and to processes of using the same. More particularly, the invention relates to novel compositions having antibacterial activity which are useful in the treatment of bacterial infections, especially those caused by drug-resistant strains of pathogenic micro-organisms. The invention also includes methods of treatment of h mans and animals infected with those strains of bacteria which are resistant to the usual chemotherapeutic agents.

The discovery of the in vivo anti-bacterial activity of sulfanilamide and the subsequent development of many other sulfonarnides having superior pharmacological chemotherapeutic properties has been one of the most inn por-tant advances in mans intense struggle. against infectious diseases caused by pathogenic and potentially pathogenic micro-orgamsms. The further discovery of penicillin, the isolation of many other antibiotic substances and the development of the nitrofurans has further enlarged the armamentarium of the practicing physician, so that there has now become available a host of useful chemotherapeutic agents. Some or" these are active against relatively few pathogens, whereas others are active against a broad spectrum of disease-producing micro-organisms of both Gram positive and Gram negative types.

Unfortunately, as the use of these chemotherapeutic agents became more widespread, there slowly began to appear strains of pathogenic microorganisms which were resistant to specific chemotherapeutic agents which formerly were highly active in controlling growth of the species. Originally gonococc-i, staphylococci and hemolytic streptococci were very susceptible to the sulfonarnides, but now many strains of these micro-organisms are no longer susceptible to treatment with these drugs. Similarly, there are now many strains of staphylococci which no longer yie d to treatment with penicillin. As a result of the extensive use of the streptomycin-s, many resistant strains have developed not only among the Gram negative bacilli, such as Aerohaczcr acrogenes, Psezzdozrzorzas aeruginosa and Proteus vul'garis, but also among the Gram positive cocci, particularly Streptococcus fecalis, Staphylococcus aureus and many others.

No wide spectrum antibiotic which may be used in vivo in the treatment of disease is known with which drug resistance is not encountered. Similarly, most all of the pathogenic bacteria will develop drug-re stant strains under suitable conditions. Accordingly, drug resistance is apt to be found anywhere at any time with any disease or with any chemotherapeutic agent. Drug resistance has been found to be commonplace in many hospitals under most carefully controlled san tary conditions, and even whole communities have been afflicted with onsiaughts of drug-re tant strains of infectious ro-organisms.

Resistance of pathogens to chemotherapeutic agents has become an increasingly serious problem in the modern practice of medicine. Simple diagnosis of the disease and identification of the causative organism is now not enough to determine the kind of chemotherapeutic agent to be used in the treatment. it is also necessary to determine whether or not the particular organism involved in the in fection is resistant against the drug that would ordinarily be used. Obviously this complicates the treatment of the patient and entails delay in administering elfective therapy during which the condition of the patient may worsen.

The mechanism by which drug resistance develops is not entirely clear. It has been supposed that in a random bacterial population there naturally occur strains or mutents of bacterial species which are resistant to the chemotherapeutic agent eiiec-tive against the normal bacterial population. As the drug is applied, those bacteria that are susceptible to it become depleted in number, Whereas the drug-resistant members of the population continue their growth and eventually control the disease process. Just why some bacterial bodies are able to thrive in the presence of the chemotherapeutic cordingly, there is no means now known of preventing the development of the phenomena. As a matter of fact, the

mechanism by which most chemotherapeutic agents exert their eilect upon the pathogen is unknown, and, accordingly, means of preventing the development of drug-resist ant strains of organisms and means of treating these resistant strains is largely empirical at the present time.

The present invention is based upon my discovery that drug-resistant strains of pathogenic micro-organisms can e controlled with appropriate chemotherapeuticagents just as effectively as the non-resistant strains of these same micro-organisms when there is administered with them a small amount of a pharmacologically acceptable mercury compound. By the term pharmacologically acceptable I mean to include those organic and inorganic meronly compounds which can or human patient without harm resulting chemical reactivity of the compound with the patient while at the same time introducing into the circulatory system a small but effective amount of mercury. Many such mercury compounds are known and are used in medicine. Of course, some of these are highly toxic, but may, nevertheless, be safely administered if the amount of the mercury is not too large. Fortunately, very small amounts of mercury are effective in potentiating or making effective known chemotherapeutic agents against pathogenic microorganisms in accordance with the present invention.

When the primary cause of the infection is in the urinary system, the amount of mercury compound admin-is tcred along with the chemotherapeutic agent preferably should be enough to produce detectable amounts of mercury in the urine. Naturally, the amount of mercury con pound administered to accomplish this purpose Will vary somewhat with the nature of the mercurial drug, but this is easily determined by simple laboratory tests.

As one example of the invention an actual case history will be related. A twenty-six year old white female who had acute follicular face had been treated with penicillin and tetracycline by usual accepted procedures for one week without improvement after which acetyl sulfisoxazole (N -(acetyl- 3,4-dimethyl-5-isoxazolyl)sulfanilamide, sold under the trade name Gantrasin Acetyl), was administered at the rate of one gram every six hours for one week with no apparent improvement in the condition of the patient. Following this, the patient was treated orally with a combination of one gram of acetylsulfoxazole and 18 mg. of chlormerodrin (sold under the trade name Neohydrin) every six hours. After four days of treatment there was definite clearing of the tonsillitis and the carbuncle.

Many more such case histories could be related. For example, a seventy-four year old white from post-operative cystitis due to beta-hemolytic streptococcus was treated with chloramphenicol toin for two weeks with no improvement. When mercurophylline sodium was administered orally along with chloramphenical and nitrofurantoin at the rate of 3. mg.-

of the mercury compound four times daily, the urine was found to be negative after five days of treatment.

Instill another case, a sixty-five year old white female agent is not known, and, ac-

be administered to an animal from toxicity or 1 tonsillitis and carbuncle of the male sufferingand nitrofuransewers suffering with cystitis and pyelitis was found to be harboring a culture of Bacillus roteus which was resistant to all drugs. This patient had been on oxytetracycline for two weeks with no ellect. She was then treated with one-half gram of N -(acetyl-3,4-dimethyl-S-isoxazolyl)- sultanilamide and 9 mg. of chlormerodrin orally every six hours. Her symptoms improved in twenty-four hours, and the urine was negative in seventy-two hours.

A substantial number of other conditions due to drug resistant strains of pathogenic micro-organisms have been successfully treated with chemotherapeutic agents in combination with mercury compounds. Among these may be mentioned post-operative pyelitis and cystitis due to beta-hemolytic streptococcus, hemorrhagic cystitis in which E. coli was identified, cystitis due to B. coli and B. proteus (sensitive only to neomycin), cystitis due to Staph. albus, and still others. The causative'agent was resistant in. each case to commonly used therapeutic agents including penicillin, nitrofurantoin, oxytctracycline, chlortetracycline, tetracycline, chloramphenical, acetyl sulfisoxazole and other normally efiective sulfa drugs, antibiotics and nitrofuran derivatives.

The pharmacologically acceptable mercury compounds that may be used in combination with sulfonamides, antibiotics, nitrofurans and other chemotherapeutic agents are as stated above those which may be taken preferably by the oral route without undue toxic reaction. The mercury compounds for this purpose are usually complex compounds, but many are sold under various trivial and tradenames. Among the suitable mercury compounds for use in practicing the present invention are disodium N-[3-(carboxymethylrnercaptomercuri)-2- methoxypropyl] -ot-camphoramate,

8- 2'-m ethoxy-3 '-hydroxymercuripropyl) -coumarin- 3-carboxylic acid,

B-methoxy-'y-hydroxymercuripropylamide of trimethylcyclopentanedicarboxylic acid,

dehydro-2- [N- 3 '-hydroxymercuri-2'-rnethoxyethoxy)propylcarbamyl] phenoxyacetic acid,

sodium o-[ (3-hydroxymercuri-Z-methoxypropyl) carbamyl]phenoxyacetate,

sodium 8-(2'-methoxy-3'-hydroxymercuripropyl)- coumarin-B-carboxylate,

[3-(chloromercuri) -2-methoxypr0pyl]urea,

sodium 1- (3 -hydroxymercuri-2-methoxypropyl) '-3 succinylurea These are more generally known under the names Mercaptomerin sodium, Mercumatilin, Mercurophylline, Merethoxylline, Mercurin, Neohydrin (Chlormerodrin, N.N.R.), and Meralluride, U.S.P. Also available are mercurialswhich are sold under the name, Merbaphen, Mersalyl U.S.P., Meragidone, Merethoxylline, Diurgin and still others which may also be used in practicing the present invention.

Most of the above mercury compounds are used in medicine as diuretic agents at various dosage levels. The mercury compound used in practicing the present invention is not used as a diuretic and hence may be used in considerably smaller amounts than the amount ordinarily used to induce diuresis. However, there may be conditions where the full amount of mercury as prescribed as a diuretic may be used in combination with an anti-bacterial agent in practicing my invention.

In many cases, as-little as one milligram of mercury per day in the form of one of the above named mercury compounds may be used effectively in practicing the present invention. Larger amounts, of course, may beused so ,that in general the present invention may be said to contemplate the use of from 1-150 mg. of mercury per day. for an adult. Thismay take the form of capsules or tabletswhich may be administered from 1 to 10 times per day. Thus the unitdosages of mercury may vary from A mg. up to about 150, mg.

As a specific instance, tablets containing an antibiotic, a sulfonamide or a therapeutically etfective-nitrofuran derivative and containing also 18.3 mg. of chlormerodrin (equivalent to It) mg. of metallic mercury) may be administered from one to ten times per day. The maximum dosage of mercury will, of course, depend upon the toxicity of the particular mercury compound used and fortunately these limits have been well defined by medical experience.

The amount of anti-bacterial agent in the composition will generally be the same as that ordinarily prescribed for the condition under treatment but in some instances may be less. For instance, the most common dosage level of the tetracyclines is 250 mg. of the antibiotic taken four times daily. This is a purely arbitrary level and may be increased in some'situatious or decreased to as low as mg. taken four times daily. Of course, in the treatment of children and small animals still smaller amounts of the antibiotic may be used.

From this it will be seen that a suitable dosage unit of a preferred composition of the present invention would include 3-10 mg. of mercury in the form of one of the above-identified mercurial compounds and 250 mg. of a tetracycline antibiotic. More specifically, a dosage unit might contain 18.3 mg. of chlormerodrin and 250 mg. of tetracycline with any desired pharmaceutically acceptable carrier. These units may directed by the attending physician.

Treatment of a wide variety of diseased conditions caused by difierent kinds of pathogens with the new compositions of the present invention has shown that the potentiating, or enhancing effects of the mercurial compounds is not related to the nature of the specific antibacterial agent that is used. It appears to be more a function of the action of the mercury on some metabolic process in the micro-organism rather than on the antibacterial agent itself. Thus, the invention is not particularly concerned with any specific anti-bacterial agent. Antibiotics such as chloramphenicol, penicillin and its analogs, tetracycline, chlortetracycline, oxytetracycline, demethyltetracycline, erythromycin, streptomycin, oleandomycin, kanamyciu, novobiocin, vancomycin and still others may be used in combination with mercury compounds of the type disclosed hereinaoove against pathogenie micro-organisms which are resistant to these anti-v biotics.

Similarly, various sulfonamides and mixtures thereof may be made active against sulfonamide-resistant pathogens by administering concurrently mercury compounds of the type mentioned. Among the various sulfonamides that may be employed with mercury compounds in practicing the present invention are sulfadiazine, sulfamethazine,.sulfamerazine, sulfisomidine, phthalylsulfathiozole, sulfodimethoxine, sulfamethoxypyridazine, succinylsulfathiazole, sulfisoxazole, thiazolsulfone and others.

Likewise, the more recently available nitrofuran derivatives such as nitroturantoin and nitrofurazone may be used with the above named mercury compounds in preparing compounds for practicing the invention.

As noted above, these anti-bacterial agents will usually be used in the same amounts in treating conditions caused by drug-fast micro-organisms as they would in infections wherein the micro-organisms are sensitive to the particular anti-bacterial agent employed. Likewise, the frequency of the dosage schedule will remain unchanged. However, because of the increased activity of the anti-bacterial agents due to the coaction of the mercury compound, the length of treatment may be decreased in many instances.

As noted before, practically all disease-producing microorganisms may become resistant to drugs under appropriate conditions. Fortunately, the use of mercury compounds With anti-bacterial agents as described hercinabove makes the latter eifective against those strains of microorganisms which have become resistant so that it is possible to use the composition of the present invention against annost any gram positive or gram negative pathogen in conditions wheresome or all of the invading micro-organisms are drug-resistant.

be taken 4 to 6 times per day as.

For example, the new compositions of the present invention can be used in the treatment of pyelitis and cystitis caused by B. hemolytic streptococcus, in cystitis caused by B. proteus and E. coli and other micro-organisms, in treating ostecmyelitis, mastitis, boils, abcesses and acne caused by S. aurezts and S. albus, in the treatment of pneumonia, sinusitis, otitis media, peritonitis and a variety of other infections caused by Szr. pyogene's, Sir. pneumococci and other streptococci, in various inflammatory and suppurative processes caused by B. vulgarz's, in the treatment of dysentery caused by Slzig. dysentariae, in the treatment of sinusitis, otitis, meningitis caused by H. influenzae, gastritis caused .by V. phosphorescent", diarrhea caused by P. aerugz'nosa and other diseases caused by micro-organisms of the groups Fusiformis, Corynebacterium, Mycobacterium, Salmonella, and other pathogens. Of course, infections caused by these particular micro-organisms will normally be treated by the drug of choice effective against them au mented, however, with the mercury compoundto ofiset the drug-fastness of the invading organism.

Example 1 Five hundred parts by weight of sulfisoxazole, 50 parts by weight of lactose and 50 parts by Weight of corn starch are micropulverized and granulated with a ten percent gelatin solution in a Day mixer. The granulation is passed through a 6 mesh screen while Wet, spread on trays and dried overnight at 100 F. The granulation is thenscreened through a 12 mesh screen and mixed with 15 parts by Weight of tale, parts by weight of magnesium stearate and 9.15 parts by Weight of chlorrnerodrin. The powdered mixture is then pressed on a single punch rotary tabletting machine to tablets of a desired size.

One or two tablets containing 500 mg. of suliisoxazole and 9.15 mg. of chlormerodrin may be taken 46 times daily for the treatment of furunculosis, cellulitis, pyleomephritis, cystitis and other conditions.

Example I! Five hundred parts by weight of sulfadiazine, 11 parts by weight of a six percent solution of methyl cellulose and25 parts by weight of corn starch as a 16 percent paste is granulated, passed through a 6 mesh screen and dried at 110 F. The granulation is then ground and passed through a 12. mesh screen, thoroughly mixed with 9.15 parts by weight of chlormerodrin, 15 parts by Weight of tale, 5 parts by Weight of magnesium stearate, 15 parts by weight of corn starch and 15 parts by weight of sodium alginate. The powdered mixture is then formed into ta lets of a desired size by conventional tablet making procedures.

Gne or two of the above tablets containing 500 mg. of suliadiazine and 9.15 of chlormerodrin are used in the treatment of conditions caused by sulfadiazine sensitive and resistant micro-organisms.

Example III Example IV Two hundred and fifty mg. of tetracycline and 7.5 mg. of chlormerodrin are filled into hard shell gelatin capsules. One or two capsules every four to six hours provide ellective therapy for the treatment of cellulitis, the pneumonias and urinary tract infections.

Example V tablets containing mg. of sodium 250 mg. of chloramphenicol and salt of p-methoxy-y-hydroxymer- Tablets containing 50 mg. of nitrofurantoin and 5 mg. of 8-(2'-methoxy 3 hydroxymercuripropyl)-coumarin- 3-carboxyiic acid (mercumallylic acid) taken orally four to times daily are eifective in the treatment of bacteremias and urinary infections.

Example VII Tablets containing 500 of kanarnycin and 10 mg. of calomel are useful in p-re-operative bacterial sterilization and for bacterial diseases of the bowels such as typhoid and para-typhoid.

I claim:

1. A method of controlling systemic infections in which pathogenic microorganisms resistant to specific antibacterial sulfon-amides are present which comprises administering orally to the infected animal a therapeutically effective amount of a sulfonamide efiective against nonresistant strains of the infecting microorganism and a subdiuretic amount of, a physiologically acceptable r-ganic mercury compound in amounts sufiicient to affect the metabolic process responsible for the sultom amide resistance of the microorganism.

2. A method in accordance with claim 1 in which 50 to 500 milligrams of sulfisoxazole and 1 to 50 milligrams of chlormerodrin are administered orally.

3. A -ethod of controlling systemic infections in which pathogenic microorganisms resistant to specific antibacterial antibiotics are present which comprises administering orally to the infected animal a therapeutically effective amount of an antibiotic effective against nonresistant strains of the infecting microorganism and a subdiuretic amount of a physiologically acceptable organic mercury compound in amounts suflicient to affect the metabolic process responsible for the antibiotic resistance of the microorganism.

4. A method in accordance with claim 3 in which 50 to 250 milligrams of tetracycline and to milligrams of chlormerodrin are administered orally.

5. A method in accordance with claim 3 in which 50 to 250 milligrams of chloramphenicol and to 150 milligrams of chlormerodrin are administered orally.

6. A method of controlling systemic infections in which pathogenic microorganisms resistant to specific antibacterial nitrofurans are present which comprises administering orally to the infected animal a therapeutically eltective amount of a nitrofuran effective against nonresistant strains of the infecting microorganism and a subdiuretic amount of a physiologically acceptable organic mercury compound in amounts suflicient to affect the metabolic process responsible for the nitroiuran resistance of the microorganism.

'7. A method in accordance with claim 6 in which 10 to 169 milligrams of nitrofurantoin and 1 to 150 milligrams of mercurnallylic acid are administered orally.

8. A composition of matter for the systemic treatment of infections in which pathogenic microorganisms resistant to specific sulfonamides are present which comprises a pero-ral dosage unit of a dry mixture in which the therapeutic ingredients consist essentially of a therapeutically eifective amount of a physiologically acceptable sulfonamide to which nonresistant strains of said pathogenic microorganisms are susceptible and a subdiuretic quantity of a physiologically acceptable organic mercury compounds in an amount sufficient to afiect the metabolic process responsible for the drug resistance of the microorganism.

9. A composition of matter in accordance with claim 8 in which the therapeutic ingredients are 50 to 500 milligrams of suliodiazine and A to 150 milligrams of chloromerodrin.

10. A composition of matter in accordance with claim 8 in which the therapeutic ingredients are 50 to 500 milligrams of sulfisoxazole and V to 150 milligrams of chlormerodrin.

11. A composition of matter for the systemic treatment of infections in which pathogenic microorganisms resistant to specific antibiotics are present which comprises a peroral dosage unit of a dry mixture in which the therapeutic ingredients consist essentially of a therapeutically effective amount of a physiologically acceptable antibiotic to which nonresistant strains of said pathogenic microorganisms are susceptible and a subdiuretic quantity of a physiologically acceptable organic mercury compound in an amount sufficient to affect the metabolic process responsible for the drug resistance of the microorganism.

12. A composition of matter in accordance with claim 11 in which the therapeutically effective ingredients are 50 to 250 milligrams of tetracycline and to .150 milligrams of chlormerodrin.

13. A composition of matter in accordance with claim 11 in which the therapeutically effective ingredients are 50 to 250 milligrams of chloramphenicol and A to 150 milligrams of chlormcrodrin.

14. A composition of matter in accordance with claim 11 in which the therapeutically effective ingredients are 50 to 250 milligrams of demethyltetracycline and A to 150 milligrams of chlormerodrin.

15. A composition of matter in accordance with claim 11 in which the mercury compound is chlormerodrin.

16. A composition of matter in accordance with claim 11 in which the mercury compound is the sodium salt oi B-methoxydicarboxylic acid.

17. A composition of matter in accordance with claim 11 in which the mercury compound is sodium 1-(3-hydroxymercuri-Z-methoxypropyl) -3-succinylurea.

18. A composition of matter for the systemic treatment of infections in which pathogenic microorganisms resistant to specific nitrofurans are present which comprises a peroral dosage unit of a dry mixture in which the therapeutic ingredients consist essentially of a therapeutically effective amount of a physiologically acceptable nitrofuran to which nonresistant strains of said pathogenic microorganismsare susceptible and a subdiuretic quantity of a physiologically acceptable .organic mercury compound in an amount sufficient to aiiect the metabolic process responsible for the drug resistance of the microorganism.

19. A composition of matter in accordance with claim 18 in which the therapeutically efiective ingredients are 10 to milligrams of nitroiurantoin and l to milligrams of mercumallylic acid.

References Cited in the file of this patent UNlTED STATES PATENTS 1,760,000 Reddy May 27, 1930 1,919,729 Kharasch July 25, 1933 1,919,732 Kharasch July 25, 1933 2,014,676 Weed Sept. 17, 1935 2,070,080 Hart Feb. 9, 1937 2,206,804 Burtner -a -a July 2, 1940 1 2,206,805 Burtner July 2, 1940 2,505,681 Martin et al. .2 Apr.-25, 1950 2,776,970 Lintner" Jan. 8, 1957 2,830,011 Parker et al. -2 Apr. 8, 1958 2,831,878 Ensminger et a1 Apr. 22, 1958 2,880,130 Johnson --.-'-*a Mar. 31, 1959 2,880,138 Johnson Mar. 31, 1959 2,899,432 Rudner .t -n Aug. 11, 1959 2,927,052 Moudry Mar. 1, 1960 2,964,447 Hosmer Dec. 13, 1960 FOREIGN PATENTS 759,577 Great Britain Oct. 17, 1956 784,659 Great Britain Oct. 16, 1957 OTHER REFERENCES Chem. Abstracts 41, ii-3440c; #4l18d (1947).

Chem. Abstracts 43, #P1805c; #9153b (1949).

Chem. Abstracts 44, #P7030a; #144021; #795b (1950).

Chem. Abstracts 45, #26290, #PS 186b, #980311 (1951).

Chem. Abstracts 46, t-"2517i, f; #5199g (1952).

Chem. Abstracts 47, #P8776i; #P12423d; -#P12425g (1953).

Chem. Abstracts 48, #5350h; #54351? (1954).

Chem. Abstracts 49, -#P3484b; #775113; #4168c (1955).

Chem. Abstracts 50, #8139f (1956).

Chem. Abstracts 51, #90931 (1957)..

Chem. Abstracts 52, #14977a;'#18991b (1958).

Chem. Abstracts 54, #11142e (1960). 

3. A METHOD OF CONTROLLING SYSTEMIC INFECTIONS IN WHICH PATHOGENIC MICOROORGANISMS RESISTANT TO SPECIFIC ANTIBATERIAL ANTIBIOTICS ARE PRESNET WHICH COMPRISES ADMINOSTERING ORALLY TO THE INFECTED ANIMAL A THERAPEUTICALLY EFFECTIVE AMOUNT OF AN ANTIBIOTIC EFFECTIVE AGAINST NONRESISTANT STRAINS OF THE INFECTING MICROORGANISM AND A SUBDIURETIC AMOUNT OF A PHYSIOLOGICALLY ACCEPTABLE ORGANIC MERCURY COMPOUND IN AMOUNTS SUFFICIENT TO AFFECT THE METABOLOC PROCESS RESPONSIBLE FOR THE ANTIBIOTIC RESISTANCE OF THE MICROORGANISM. 